Objective To investigate the characteristics of human immunodeficiency virus(HIV-1)V3 loop amino acid mutations among AIDS patients in Shenzhen. Methods Fragments of HIV-1 env gene were amplified by RT-nested-polymerase chain reaction(PCR)from plasma of the AIDS patients in Shenzhen.The PCR products were then directly sequenced and the sequences were aligned,translated and analyzed. Results Sequence analysis showed that there were B and CRF01-AE HIV-1 subtypes in Shenzhen.The V3 loop terra peptide GPGQ accounted for about 48%,GPGR about 36%,other V3 loop terra peptide forms were 16%;the V3 loop central motif in the majority(76.9%)of the CRF01-AE strains was GPGQ,the majority(75%)of the B strains was GPGR.We also found some unusual terra peptide compositions:DQDR and DQGQ on the tip of V3 loop,some amino acid mutations at specific of V3 loop associated with SI phenotype were very common in AIDS patients(80%). Conclusions The V3 loop amino acid mutations were very common in AIDS patients,the V3 loop central motif in the majority was GPGQ and GPGR in Shenzhen,the mutations shows the change of HIV-1 phenotype and predicts that disease progress to AIDS.

In order to construct generally efficient cell immortalization vector, pTP-hTERT, we modified the traditional piggyBac (PB) transposon using artificial synthesis, PCR and enzyme digestion. The modified vector contained the necessary transposon elements, a PB transposase expression cassette, a co-expression selectable element and a human telomerase reverse transcriptase (hTERT) expression cassette. The co-expression selectable element had two markers, enhanced green fluorescent protein (EGFP) gene and puromycin-resistance (Puro) gene, linked by porcine teschovirus-1 2A peptide (P2A). To validate the functionality of vector elements, we transfected pTP-hTERT into HEK293 cell, selected the positive cell clones and then conducted RT-PCR, Western blotting (WB) and Tail-PCR, methylene blue staining and statistic analysis on selected cells. The results of sequencing and cell culture show that the pTP-hTERT was constructed successfully and the positive cell could be selected by puromycin. The WB results, P2A cutting EGFP and Puro fusion protein with high efficiency, reflected the selectable element worked. The sequencing result of Tail-PCR confirmed the vector integrated into the genome through transposition. The results of methylene blue staining and statistic analysis indicated the clone of positive cells triggered by pTP-hTERT significantly increased (P < 0.01) compared with control group. The construction of pTP-hTERT provides an efficient tool for establishing immortalized cell lines and a demonstration for building other eukaryotic plasmids.
Cell Line ; DNA Transposable Elements ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Polymerase Chain Reaction ; Telomerase ; genetics ; Transfection

The construction of a network resource data base for clinical skill teaching aiming at sharing excellent teaching resources was completed by integrating all kinds of teaching resource including characters,eourseware,pictures,cartoons,videos and examination questions,etc.In view of the generally weak situation of the current domestic education resource data base construction,the contents,guiding ideology,principle,object,orientation,function and the management mechanism of the network resource data base construction were devised for clinical skills teaching.Suggestions for specific ideas and construction problems were proposed to promote the construction and optimization of the network resource database for clinical skills teaching.

OBJECTIVE: To determine the feasibility whether the bovine jugular venous conduit (BJVC) can be fixed with polyepoxy compound (PC). METHODS: Twenty-four BJVCs were divided into 3 groups and fixed with polyepoxy compound (PC group, n = 8), glutaraldehyde (GA group, n = 8), and unfixed group (Control group, n = 8), respectively. The morphologic and mechanical properties of BJVCs in the 3 groups, including thickness, diameter, moisture content, denaturation temperature, tensile strength, elongation at break, and fixation index were measured. The rat subcutaneous model for the assessment of tissue calcification was used. The calcium content in bovine jugular vein patches and valves was determined by flame atomic absorption spectrophotometer. RESULTS: There was no difference in the wall thickness, diameter, and tissue water content between PC and the control group, but significant difference was found between GA and PC groups. The mechanical properties of PC group and GA group were not significantly different, but they were better than those of the control group. GA-fixed BJVC samples showed clear calcification, while PC fixed BJVC were calcified significantly less. CONCLUSION PC is an effective and suitable choice for the treatment of BJVC since it can effectively preserve the structure and the anti-reflow function of valves in bovine jugular vein and it has better anti-calcification properties.
Animals ; Biocompatible Materials ; Bioprosthesis ; Blood Vessel Prosthesis ; Cattle ; Cross-Linking Reagents ; pharmacology ; Epoxy Compounds ; pharmacology ; Jugular Veins ; Polymers

This study aims to analyse the value of CODEHOP RT-PCR in the detection of Flavivirus. According to the amino acid sequences of polyproteins of different flaviviruses published in GenBank, a pair of primers was designed using the CODEHOP method. One-step RT-PCR was used to detect Japanese encephalitis virus strain JEV1201, Dengue virus strain JKD001, and yellow fever virus vaccine YV6161. BLAST analysis and phylogenetic analysis were performed after the RT-PCR products of nucleocapsid genes were sequenced. The results showed that this method could amplify Flavivirus specifically, and the size and sequence of the target fragment accorded with the anticipated result. JEV1201 had the highest homology to Japanese encephalitis virus strain YL2009-4/YC2009-3, belonging to the branch of the phylogenetic tree of Japanese encephalitis virus strains. JKD001 had the highest homology to Dengue virus strain DENV-2/ID/1022DN/1975, belonging to the branch of the phylogenetic tree of Dengue virus strains. YV6161 had the highest homology to Yellow fever virus strain 17D, belonging to the branch of the phylogenetic tree of Yellow fever virus strains. In conclusion, the method of CODEHOP RT-PCR can be effectively used to detect, identify, and phylogenetically analyse Flavivirus.
DNA Primers ; genetics ; Flavivirus ; classification ; genetics ; isolation & purification ; Flavivirus Infections ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Nonstructural Proteins ; genetics

Objective To approach potential abnormalities of neuro-chemical compounds in cerebrum of obsessive-compulsive disorder and to analyze the relationship among the abnormalities with duration of illness and symptom severity. Methods 19 patients with obsessive-compulsive disorder and 22 comparison subjects were scanned by 1H magnetic resonance spectroscopy to exam bilateral prefrontal lobe, temporal lobe, parietal lobe, occipital lobe, anterior cingutate and caudate nucleus, hippocampus as well as splenium and genu of corpus callosum,thalamus. The ratios of N-acetylaspartate (NAA) and choline (Cho) to creatine (Cr) were respectively recorded.The severities of obsessive-compulsive symptoms were assessed by Yale-Brown Obsessive-Compulsive Scale. Results ( 1 ) The ratios of NAA/Cr in right caudate nucleus and left hippocampus were higher than those in normal controls (1.23 ±0.37 vs 1.0 ±0.33, t=2.14, P=0.039;1.34 ±0.25 vs 1. 10 ±0.45, t=2.41, P=0.021 ), but NAA/Cr ratios in genu of corpus callosum were lower(0. 97 ±0.30 vs 1.19 ± 0. 26, t = -2.47, P= 0.018 ).(2) The Cho/Cr ratios in right caudate nucleus and left temporal lobe were higher than those in control groups (1.09±0.51 vs0. 67±0.21,t=3.32, P=0.003;2.58±0.62vs0.84±0.17, t=2.21,P=0.03). (3) The duration of illness was negatively correlated with the Cho/Cr ratios of right prefrontal lobe ( r = - 0. 507, P =0. 027 ). (4) The NAA/Cr ratios of genu of corpus callosum as well as the Cho/Cr ratios of left temporal lobe were negatively correlated with YBOCS score ( r 1 = - 0.457, P = 0.049; r 2 = - 0. 585, P = 0.009 ). Conclusion Neuronal functions increase in right caudate nucleus and left hippocampus which are opposite in genu of corpus callosum. These abnormalities may play an important role in pathogenesis of OCD.

AIM: To investigate the effects of Yiqi Huoxue prescription and Bushen Shengsui prescription on the expression of Notch-1 and Jagged1 protein in ischemic penumbra after local cerebral ischemia/reperfusion in intraluminal thread occlusion of the middle cerebral artery (MCAO) rats. METHODS: The local cerebral ischemia/reperfusion model was established by intraluminal thread occlusion of the middle cerebral artery. The animals were randomly divided into pseudo surgery group (sham group), model (MCAO) group, Yiqi Huoxue prescription (MCAO+Yiqi Huoxue) and Bushen Shengsui prescription (MCAO+Bushen Shengsui) groups. Using the techniques of immuno-histochemical staining, the expression of Notch-1 and Jagged1 was observed at 1, 2 and 3 weeks in ischemic penumbra of the cortex of frontal and parietal lobe. RESULTS: A value and positive unit of Notch-1 and Jagged1 protein expression in model group were higher than those in sham in ischemic penumbra of the cortex of frontal or parietal lobe after local cerebral ischemia/reperfusion in MCAO rats (P<0.05 or P<0.01). A value and/or positive unit of the expression in Yiqi Huoxue prescription and Bushen Shengsui prescription groups were lower than those in model group (P<0.05 or P<0.01). CONCLUSION: Yiqi Huoxue prescription and Bushen Shengsui prescription affect the proliferation and differentiation of neural stem cells, depress Notch signal transduction after local cerebral ischemia/reperfusion in MCAO rats by inhibiting the expression of Notch-1and Jagged1 protein.

Objective To discuss the diagnosis and treatment of cerebral sparganosis. Methods To summary four cases of cerebral sparganosis, focusing on the clinical course and imaging findings, with the goal of better diagnostic skills. Results All 4 cases had some kind of misdiagnosis and improvement after surgery or parasiticidal pharmacotherapy. Cerebral MRI and CT scans revealed relatively extensive white matter degeneration and focal enhancements. Subsequent scans showed changes in shape and location of the enhanced foci, indicating the migration of sparganum. Pathologic findings of 3 patients who had undergone surgery showed granuloma and sparganum. Conclusions Cerebral sparganosis has relatively special manifestions on imaging, which are of diagnostic value. The spaganum should be as completely removed as possible during surgery.

OBJECTIVETo study immunomodulating activity of Lonicera Japonica flavone by investigating immune enzymatic activity of serum and antoxidized activity of lymphoid organs in mice.

METHODSFifty KM mice were randomly divided into control group, model group, low dose group, middle dose group and high dose group(n = 10), respectively. And low dose group, middle dose group and high dose group were given Lonicera Japonica flavone with 100 mg/kg, 200 mg/kg and 400 mg/kg every day, respectively, while control group and model group were administered with NS. After continuously giving drug 7 weeks, other groups were injected with Dexamethasome (Dex: 25 mg /kg) for 3 days by subcutaneous injection, but the control group were treated with NS. And after giving Lonicera Japonica flavone 1 week simultaneously, organ indexes , the activity of acid phosphatase (ACP), alkaline phosphatase (AKP) and lysozyme (LSZ) in serum , and the content of monoamine oxidase (MAO), total antioxidant capacity (T-AOC), total superoxide dismutase (SOD) and malondialdehyde (MDA) in lymphoid organs in mice were tested, respectively.

RESULTSLonicera Japonica flavone could significantly improve the organ indexes, and significantly improve the activity of ACP, AKP and LSZ in serum, and significantly improve the contents of T-AOC and SOD, but reduce that of MAO and MDA in lymphoid organs in immunosuppressed mice.

CONCLUSIONIonicera Japonica flavone can significantly improve the activity of immune enzyme in serum and the antioxidized activity of lymphoid organs in mice. It suggests that Ionicera Japonica flavone has a good immunomodulatory effects.

Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Antioxidants ; metabolism ; Flavones ; pharmacology ; Immunomodulation ; Lonicera ; chemistry ; Malondialdehyde ; metabolism ; Mice ; Monoamine Oxidase ; metabolism ; Muramidase ; blood ; Superoxide Dismutase ; metabolism

As a common computer keyboard and a mouse can't meet the requirements of some special occasions, this paper introduces the design of a controlling switch set based on RS-232 for capturing the image at the medical image workstation. Its working principles, circuit, assembly together with its associated software controller are given in detail. The control switch set features easy manufacturing, low cost and a good reliability.
Equipment Design ; Image Processing, Computer-Assisted ; instrumentation ; methods ; Software Design